SASDAQ4: 6:3 complex of GAD:CaM (Calmodulin + Glutamate decarboxylase, GAD)

基本情報

• 登録情報: データベース: SASBDB / ID: SASDAQ4

試料

6:3 complex of GAD:CaM

• Calmodulin (protein), Homo sapiens
• Glutamate decarboxylase (protein), GAD, Petunia x hybrida

機能・相同性

分子機能:

glutamate decarboxylase / glutamate decarboxylase activity / : / glutamate catabolic process / establishment of protein localization to mitochondrial membrane / type 3 metabotropic glutamate receptor binding / regulation of synaptic vesicle endocytosis / negative regulation of high voltage-gated calcium channel activity / positive regulation of cyclic-nucleotide phosphodiesterase activity / negative regulation of calcium ion export across plasma membrane / regulation of synaptic vesicle exocytosis / regulation of cardiac muscle cell action potential / response to corticosterone / nitric-oxide synthase binding / positive regulation of ryanodine-sensitive calcium-release channel activity / regulation of cell communication by electrical coupling involved in cardiac conduction / negative regulation of peptidyl-threonine phosphorylation / protein phosphatase activator activity / positive regulation of phosphoprotein phosphatase activity / adenylate cyclase binding / catalytic complex / detection of calcium ion / regulation of cardiac muscle contraction / negative regulation of ryanodine-sensitive calcium-release channel activity / positive regulation of DNA binding / regulation of cardiac muscle contraction by regulation of the release of sequestered calcium ion / phosphatidylinositol 3-kinase binding / regulation of release of sequestered calcium ion into cytosol by sarcoplasmic reticulum / enzyme regulator activity / positive regulation of protein dephosphorylation / regulation of calcium-mediated signaling / titin binding / positive regulation of protein autophosphorylation / voltage-gated potassium channel complex / calcium channel complex / response to amphetamine / activation of adenylate cyclase activity / substantia nigra development / adenylate cyclase activator activity / nitric-oxide synthase regulator activity / regulation of heart rate / sarcomere / protein serine/threonine kinase activator activity / positive regulation of peptidyl-threonine phosphorylation / regulation of cytokinesis / positive regulation of nitric-oxide synthase activity / mitochondrial membrane / positive regulation of protein serine/threonine kinase activity / spindle microtubule / synaptic vesicle membrane / spindle pole / response to calcium ion / G2/M transition of mitotic cell cycle / calcium-dependent protein binding / disordered domain specific binding / pyridoxal phosphate binding / myelin sheath / growth cone / vesicle / transmembrane transporter binding / calmodulin binding / G protein-coupled receptor signaling pathway / protein domain specific binding / centrosome / calcium ion binding / protein kinase binding / protein-containing complex / nucleus / plasma membrane / cytoplasm / cytosol

ドメイン・相同性:

Glutamate decarboxylase / Pyridoxal phosphate-dependent decarboxylase / Pyridoxal-dependent decarboxylase conserved domain / EF-hand domain pair / Pyridoxal phosphate-dependent transferase, major domain / Pyridoxal phosphate-dependent transferase / EF-hand, calcium binding motif / EF-Hand 1, calcium-binding site / EF-hand calcium-binding domain. / EF-hand calcium-binding domain profile. / EF-hand domain / EF-hand domain pair

構成要素:

Calmodulin-3 / Glutamate decarboxylase

• 生物種: Homo sapiens (ヒト); Petunia x hybrida (ナス科)
• 引用: ジャーナル: J Mol Biol / 年: 2009; タイトル: A common structural basis for pH- and calmodulin-mediated regulation in plant glutamate decarboxylase.; 著者: Heinz Gut / Paola Dominici / Stefania Pilati / Alessandra Astegno / Maxim V Petoukhov / Dmitri I Svergun / Markus G Grütter / Guido Capitani / ; 要旨: Glutamate decarboxylase (Gad) catalyzes glutamate to gamma-aminobutyrate conversion. Plant Gad is a approximately 340 kDa hexamer, involved in development and stress response, and regulated by pH and binding of Ca(2+)/calmodulin (CaM) to the C-terminal domain. We determined the crystal structure of Arabidopsis thaliana Gad1 in its CaM-free state, obtained a low-resolution structure of the calmodulin-activated Gad complex by small-angle X-ray scattering and identified the crucial residues, in the C-terminal domain, for regulation by pH and CaM binding. CaM activates Gad1 in a unique way by relieving two C-terminal autoinhibition domains of adjacent active sites, forming a 393 kDa Gad1-CaM complex with an unusual 1:3 stoichiometry. The complex is loosely packed: thanks to the flexible linkers connecting the enzyme core with the six C-terminal regulatory domains, the CaM molecules retain considerable positional and orientational freedom with respect to Gad1. The complex thus represents a prototype for a novel CaM-target interaction mode. Thanks to its two levels of regulation, both targeting the C-terminal domain, Gad can respond flexibly to different kinds of cellular stress occurring at different pH values.

登録者

• Maxim Petoukhov (EMBL-Hamburg, European Molecular Biology Laboratory (EMBL) - Hamburg outstation, Notkestraße 85, Geb. 25A, 22607 Hamburg, Deutschland, Germany)

モデル

• モデル #67: ; タイプ: mix / ソフトウェア: Bunch / ダミー原子の半径: 1.90 A / 対称性: P6/P1; Omokage検索でこの集合体の類似形状データを探す (詳細)

試料

• 試料: 名称: 6:3 complex of GAD:CaM / Sample MW: 357.22 kDa / Entity id: 58 / 61
• バッファ: 名称: HEPES / 濃度: 50.00 mM / PK: 7 / pH: 7.5 ; コメント: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; 組成: KCl 50.000 mM

要素 #58

タイプ: protein / 記述: Calmodulin / 分子量: 16.84 / 分子数: 1 / 由来: Homo sapiens / 参照: UniProt: P62158
配列:

MADQLTEEQI AEFKEAFSLF DKDGDGTITT KELGTVMRSL GQNPTEAELQ DMINEVDADG NGTIDFPEFL TMMARKMKDT DSEEEIREAF RVFDKDGNGY ISAAELRHVM TNLGEKLTDE EVDEMIREAD IDGDGQVNYE EFVQMMTAK

要素 #61

名称: GAD / タイプ: protein / 記述: Glutamate decarboxylase / 分子量: 56.73 / 分子数: 6 / 由来: Petunia x hybrida / 参照: UniProt: Q07346
配列:

MVLSKTVSQS DVSIHSTFAS RYVRTSLPRF KMPDNSIPKE AAYQIINDEL MLDGNPRLNL ASFVTTWMEP ECDKLMMDSI NKNYVDMDEY PVTTELQNRC VNMIAHLFNA PLEDGETAVG VGTVGSSEAI MLAGLAFKRK WQNKMKAQGK PCDKPNIVTG ANVQVCWEKF ARYFEVELKE VKLSEGYYVM DPEKAVEMVD ENTICVAAIL GSTLNGEFED VKRLNDLLVE KNKETGWDTP IHVDAASGGF IAPFIYPELE WDFRLPLVKS INVSGHKYGL VYAGIGWVVW RNKDDLPDEL IFHINYLGAD QPTFTLNFSK GSSQVIAQYY QLIRLGYEGY KNVMENCQEN ASVLREGLEK TGRFNIISKE IGVPLVAFSL KDNRQHNEFE ISETLRRFGW IVPAYTMPPN AQHITVLRVV IREDFSRTLA ERLVRDIEKV LHELDTLPAR VNAKLAVAEE QAAANGSEVH KKTDSEVQLE MITAWKKFVE EKKKKTNRVC

実験情報

• ビーム: 設備名称: DORIS III X33 / 地域: Hamburg / 国: Germany / 線源: X-ray synchrotron
• 検出器: 名称: MAR 345 Image Plate

スキャン

タイトル: GAD:CaM complex / 測定日: 2004年11月9日 / 単位: 1/nm /

	Min	Max
Q	0.1557	4.483

距離分布関数 P(R)

ソフトウェア P(R): GNOM 4.6 / ポイント数: 285 /

	Min	Max
Q	0.1568	1.396
P(R) point	1	285
R	0	23.17

結果

I0 from Guinier: 87600 / Rg from Guinier: 5.85 nm / カーブのタイプ: single_conc /

	実験値	Porod
分子量	400 kDa	-
体積	-	630 nm3

/

	Min	Max
D	-	23
Guinier point	1	29